Novel whey-derived peptides with inhibitory effect against angiotensin-converting enzyme: In vitro effect and stability to gastrointestinal enzymes

Whey protein concentrate (WPC) was subjected to enzymatic hydrolysis by proteases from the flowers of Cynara cardunculus, and the resulting angiotensin-converting enzyme (ACE)-inhibitory effect was monitored. The whole WPC hydrolysate exhibited an IC50 value of 52.9 ± 2.9 μg/mL, whereas the associated peptide fraction with molecular weight below 3 kDa scored 23.6 ± 1.1 μg/mL. The latter fraction was submitted to RP-HPLC, and 6 fractions were resolved that exhibited ACE-inhibitory effects. Among the various peptides found, a total of 14 were identified via sequencing with an ion-trap mass spectrometer. Eleven of these peptides were synthesized de novo – to validate their ACE-inhibitory effect, and also to ascertain their stability when exposed to simulated gastrointestinal digestion. Among them, three novel, highly potent peptides were found, corresponding to α-lactalbumin f(16–26) – with the sequence KGYGGVSLPEW, α-lactalbumin f(97–104) with DKVGINYW, and β-lactoglobulin f(33–42) with DAQSAPLRVY; their IC50 values were as low as 0.80 ± 0.1, 25.2 ± 1.0 and 13.0 ± 1.0 μg/mL, respectively. None of them remained stable in the presence of gastrointestinal enzymes: they were partially, or even totally hydrolyzed to smaller peptides – yet the observed ACE-inhibitory effects were not severely affected for two of those peptides.

Research highlights
► Several peptides from whey proteins were released by nonconventional plant protease.
► Eleven peptides were tested for ACE-inhibitory activity, as such and after synthesis de novo.
► Novel alpha-lactalbumin f(16–26) peptide was released, with IC50 of 0.80 microgram/mL.
► simulated gastrointestinal digestion did not compromise ACE-inhibitory activity of best peptides.